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The combination of in situ hybridization of fluorescent probes onto chromosomes in solution and flow cytometry allows the quantitative identification of repeat sequences in individual chromosomes.
Software for the automated and accurate registration of multiple images of Drosophila melanogaster brain is reported. It is used to build a preliminary atlas of gene expression in the fly brain.
A combination of PCR stitching with next-generation sequencing results in Stitch-seq, a massively parallel method for interactome mapping. The approach is applied to human protein-protein interactions assayed in yeast two-hybrid screens.
Semantically typed data hypercubes (SDCubes) are described for the flexible management and organization of stored experimental data and are applied to high-content imaging data using the software package ImageRail.
Incorporation of one-dimensional imaging capability into a parallel microfluidic flow cytometer allows fast, low-resolution acquisition of images that permit classification of cells by automated analysis of preselected features.
A defined and simplified culture system for the derivation and growth of human induced pluripotent stem cells is reported. It permits increased efficiency of human reprogramming with an episomal approach. Also in this issue, Okita et al. describe methods for more efficient episomal reprogramming of human cells.
Human induced pluripotent stem cells are generated with episomal plasmid vectors at increased efficiency using non-transforming L-Myc and knockdown of p53. Also in this issue, Chen et al. report defined conditions for human cell reprogramming and culture.
Short hairpin RNAs, expressed from microRNA scaffold–containing vectors, efficiently silence gene expression in female germ cells as well as somatic cells in the fly. A genome-wide resource is being developed.
Rapid, high-resolution, label-free Fourier-transform infrared imaging of biological samples is made possible by combining multiple synchrotron beams with wide-field detection.
Analyzing the first and higher-order moments of the diffraction spot of a 4Pi fluorescence detection scheme facilitates two-color, three-dimensional super-resolution microscopy with ~6 nm axial and ~8–23 nm lateral resolution in a layer ~650 nm thick.
A miniature portable tomograph allows the imaging of awake, behaving rats with positron emission tomography while simultaneously measuring their behavior. This method for noninvasive whole-brain imaging can link studies of brain activity with particular behaviors.
RNAi screening combined with automated microscopy of Drosophila melanogaster cells reveal genetic interactions and allow the reconstruction of signaling pathways.
Light sheet microscopy using a scanned Bessel beam in combination with structured illumination or two-photon excitation reduces photobleaching and phototoxicity, improves axial resolution and allows isotropic three-dimensional imaging. The authors demonstrate performance of the method via fast volumetric subcellular imaging of several dynamic processes in single living cells.
The combination of an ultrahigh-resolution dual optical trap with a confocal microscope allowed single-fluorophore detection of labeled oligonucleotide binding and simultaneous measurement of angstrom-scale changes in DNA tether extension.
Confined photoactivation of photoactivatable mCherry using two-photon illumination with line-scanning temporal focusing in combination with three-dimensional localization algorithms allows three-dimensional super-resolution microscopy of cellular features at <50 nm lateral and <100 nm axial resolution and depths greater than 8 ÎĽm.
A microfluidic mixing device for multiple, rapid and automated single-molecule measurements permits the study of macromolecule properties under varying environmental conditions. Also in this issue, Gambin et al. present another microfluidic mixing device for rapid single-molecule measurements.
A laminar flow mixing microfluidic device enables single-molecule fluorescence resonance energy transfer (FRET) kinetic measurements with a time resolution of 0.2 ms, enabling the study of early binding-coupled folding and unfolding events of an intrinsically disordered protein, α-synuclein. Also in this issue, Kim et al. describe another microfluidic mixing device for single-molecule experiments.